背景:半胱氨酸双加氧酶1(CDO1)经常被甲基化,在包括乳腺癌(BC)在内的许多人类癌症中,其表达降低。然而,对BC中CDO1失活的功能和机制方面知之甚少,血清CDO1甲基化的诊断意义尚不清楚。
方法:我们对公开可用的数据库进行了生物信息学分析,并采用了MassARRAYEpiTYPER甲基化测序技术来鉴定BC组织与正常邻近组织(NAT)相比CDO1启动子中的差异甲基化位点。随后,我们使用针对这些CpG位点的特异性引物和探针开发了MethyLight测定法,以检测CDO1启动子的甲基化参考(PMR)的百分比。此外,基于LentiCRISPR/dCas9-Tet1CD的CDO1靶向去甲基化系统和CDO1过表达策略均用于检测BC中CDO1的功能和潜在机制。最后,评估了CDO1作为BC血清甲基化生物标志物的早期诊断价值.
结果:CDO1启动子在BC组织中高度甲基化,这与不良预后有关(p<0.05)。基于CRISPR/dCas9的靶向去甲基化系统显着降低了CDO1启动子的PMR,并增加了BC细胞中的CDO1表达。因此,这导致细胞增殖的抑制,移民和入侵。此外,我们发现CDO1通过抑制细胞周期发挥肿瘤抑制作用,促进细胞凋亡和铁凋亡。此外,我们使用MethyLight检测BC血清中的CDO1PMR,我们发现血清CDO1甲基化是早期诊断BC的有效非侵入性生物标志物。
结论:CDO1是高度甲基化的,在BC中充当肿瘤抑制基因。表观遗传编辑异常的CDO1甲基化可能在BC的临床治疗和预后中起着至关重要的作用。此外,血清CDO1甲基化有望作为早期诊断和治疗BC的有价值的生物标志物。
Cysteine dioxygenase 1 (CDO1) is frequently methylated, and its expression is decreased in many human cancers including breast cancer (BC). However, the functional and mechanistic aspects of CDO1 inactivation in BC are poorly understood, and the diagnostic significance of serum
CDO1 methylation remains unclear.
We performed bioinformatics analysis of publicly available databases and employed MassARRAY EpiTYPER methylation sequencing technology to identify differentially methylated sites in the CDO1 promoter of BC tissues compared to normal adjacent tissues (NATs). Subsequently, we developed a MethyLight assay using specific primers and probes for these CpG sites to detect the percentage of methylated reference (PMR) of the CDO1 promoter. Furthermore, both LentiCRISPR/dCas9-Tet1CD-based
CDO1-targeted demethylation system and
CDO1 overexpression strategy were utilized to detect the function and underlying mechanism of CDO1 in BC. Finally, the early diagnostic value of CDO1 as a methylation biomarker in BC serum was evaluated.
CDO1 promoter was hypermethylated in BC tissues, which was related to poor prognosis (p < .05). The CRISPR/dCas9-based targeted demethylation system significantly reduced the PMR of CDO1 promotor and increased
CDO1 expression in BC cells. Consequently, this leads to suppression of cell proliferation, migration and invasion. Additionally, we found that CDO1 exerted a tumour suppressor effect by inhibiting the cell cycle, promoting cell apoptosis and ferroptosis. Furthermore, we employed the MethyLight to detect CDO1 PMR in BC serum, and we discovered that serum
CDO1 methylation was an effective non-invasive biomarker for early diagnosis of BC.
CDO1 is hypermethylated and acts as a tumour suppressor gene in BC. Epigenetic editing of abnormal CDO1 methylation could have a crucial role in the clinical treatment and prognosis of BC. Additionally, serum CDO1 methylation holds promise as a valuable biomarker for the early diagnosis and management of BC.